FD-895 and Pladienolide B Modulate RNA Splicing Associated With Wnt Signaling Pathway

Abstract

BackgroundAlterations in RNA splicing are associated with different malignancies including leukemia, lymphoma, and solid tumors. The RNA splicing modulators such as FD-895 and pladienolide B have been investigated in different malignancies to target/modulate spliceosome for therapeutic purpose. MethodsThe different cell lines were screened using RNA splicing modulator to test in vitro cytotoxicity as well as ability to modulate RNA splicing capability via induction of intron retention (using RT-PCR and qPCR) was assessed. The Cignal Finder Reporter Array was used to screen the HeLa cell line to know the pathway affected the most by the splice modulators. Further the candidates associated with the pathways were validated at protein levels using western blot assay and gene-gene interaction study were carried out using GeneMANIA.ResultsWe show that FD-895 and pladienolide B have more apoptotic activities than conventional chemotherapy in different solid tumors. We found FD-895 or pladienolide B modulate Wnt signaling pathways and mRNA splicing. We showed that FD-895 or pladienolide B significantly down regulates not only Wnt signaling pathway-associated transcripts (GSK3β and LRP5), but also both transcript and proteins including LEF1, CCND1, LRP6, and pLRP6. ConclusionThese results indicate FD-895 and pladienolide B inhibit Wnt signaling by impeding with phosphorylation of LRP6 and modulates mRNA splicing through induction of intron retention in cervical cancer cells.